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<p>Rapid production of haploid and doubled haploid (DH) lines is essential for accelerating breeding <i>in Capsicum annuum</i>. This study evaluated the effects of carbon-based adsorbents and antioxidants on anther culture of three F₁ hybrids (Nirvin, LIAD, DiCaprio). Flower buds at the mid- to late-uninucleate stage (buds, 5–6&#xa0;mm length) were cultured on C medium containing 2&#xa0;mg L⁻¹ 2,4-D, 2&#xa0;mg L⁻¹ kinetin, 30&#xa0;g L⁻¹ sucrose, and 0.7% agar. Activated charcoal (AC), biochar (BC), and nanobiochar (NBC) were tested at three different concentrations of 0, 0.5, 1, and 2%. In the second phase, the optimal treatment was combined with ascorbic acid (AA: 20, 50, 100&#xa0;mg L⁻¹) and nano-encapsulated ascorbic acid (NEAA: 3.33, 16.66, 33.33&#xa0;mg L⁻¹). AC and BC increased embryo induction and regeneration, while NBC at 1% produced the highest embryo number (2), embryogenesis frequency (20%), and plantlets (1.46); no embryos were observed in the control. The highest regeneration (96.66%) was obtained with 1% AC. Increasing AA reduced responses, whereas 16.66&#xa0;mg L⁻¹ NEAA + 1% NBC increased embryo number (5), embryogenesis (50%), and plantlets (4.2). The highest regeneration (100%) was observed with 1% NBC + 33.33&#xa0;mg L⁻¹ NEAA. LIAD under 1% NBC + 16.66&#xa0;mg L⁻¹ NEAA showed the highest embryo number (5.6), embryogenesis (56%), and plantlets (4.5). Analysis of 30 regenerated plants with flow cytometry and chromosome counting. showed that 30–40% were haploid, and SSR analysis confirmed that the remaining plants had undergone spontaneous chromosome doubling; no mixoploid plants were observed. In total, 148 DH lines were produced.</p>

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Nanobiochar and nano-encapsulated ascorbic acid enhance androgenesis and doubled haploid production via anther culture in bell pepper (Capsicum annuum L.)

  • 期刊论文

  • 发表日期：

    2026年6月16日

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  • Mohammadvali Habibi-Silabi,
  • Jaber Panahandeh,
  • Mehran E. Shariatpanahi,
  • Alireza Motallebi Azar,
  • Mohammadreza Hassandokht

摘要

Rapid production of haploid and doubled haploid (DH) lines is essential for accelerating breeding in Capsicum annuum. This study evaluated the effects of carbon-based adsorbents and antioxidants on anther culture of three F₁ hybrids (Nirvin, LIAD, DiCaprio). Flower buds at the mid- to late-uninucleate stage (buds, 5–6 mm length) were cultured on C medium containing 2 mg L⁻¹ 2,4-D, 2 mg L⁻¹ kinetin, 30 g L⁻¹ sucrose, and 0.7% agar. Activated charcoal (AC), biochar (BC), and nanobiochar (NBC) were tested at three different concentrations of 0, 0.5, 1, and 2%. In the second phase, the optimal treatment was combined with ascorbic acid (AA: 20, 50, 100 mg L⁻¹) and nano-encapsulated ascorbic acid (NEAA: 3.33, 16.66, 33.33 mg L⁻¹). AC and BC increased embryo induction and regeneration, while NBC at 1% produced the highest embryo number (2), embryogenesis frequency (20%), and plantlets (1.46); no embryos were observed in the control. The highest regeneration (96.66%) was obtained with 1% AC. Increasing AA reduced responses, whereas 16.66 mg L⁻¹ NEAA + 1% NBC increased embryo number (5), embryogenesis (50%), and plantlets (4.2). The highest regeneration (100%) was observed with 1% NBC + 33.33 mg L⁻¹ NEAA. LIAD under 1% NBC + 16.66 mg L⁻¹ NEAA showed the highest embryo number (5.6), embryogenesis (56%), and plantlets (4.5). Analysis of 30 regenerated plants with flow cytometry and chromosome counting. showed that 30–40% were haploid, and SSR analysis confirmed that the remaining plants had undergone spontaneous chromosome doubling; no mixoploid plants were observed. In total, 148 DH lines were produced.